This applications concerns new formulations for a class of tricyclic vasopressin agonist compounds, and pharmaceutically acceptable salts thereof, as well as processes for manufacturing the formulations. The invention particularly relates to orally administered formulations of these compounds.
The art describes many methods of producing liquid or semi-solid encapsulated pharmaceutical formulations. In Bull. Tech./Gattefosse Rep. (1996), 89, 27-38, authors Shah et al. describe hard gelatin capsule technology, particularly for use in enhancing the bioavailability of poorly soluble or poorly absorbed drugs.
U.S. Pat. No. 4,620,974 (Hersh et al.) teaches a hard gelatin capsule comprising a telescoping two-piece cap with a lubricant comprising a polyethylene glycol of a molecular weight between about 200 and about 900 present in admixture with the composition at a concentration of from about 0.5 to about 25 weight percent.
WO 96/40071 (Lamberti) discloses methods and devices for producing minimal volume capsules. WO 96/41622 (Tanner et al.) teaches suspensions suitable for encapsulation in gelatin capsules, particularly including a solid phase of solid particles and a liquid phase capable of suspending the solid phase.
U.S. Pat. No. 5,641,512 (Cimiluca) teaches soft gelatin encapsulated analgesics in which the shell contains a xanthine derivative, such as caffeine.
U.S. Pat. No. 4,578,391 (Kawata et al.) describes oily compositions for antitumor agents comprising at least one sparingly oil soluble or water-soluble antitumor drug, at least one fat or oil, and at least one solubilizing adjuvant in an oily vehicle, selected from crown ether, lecithin, polyethylene glycol, propylene glycol, vitamin E, polyoxyehtylene alkylether, and sucrose esters of fatty acids.
EP 0 815 854 A1 discloses a substantially translucent, semi-solid fill material for a soft gelatin capsule, the semi-solid material being sufficiently viscous that it cannot be expelled from the capsule with a syringe at room temperature.
U.S. Pat. No. 4,744,988 (Brox) teaches soft gelatin capsules comprising a shell of gelatin, a softener and a filling of a polyethylene glycol and a low polyhydric alcohol and at least one active substance, characterized in that the shell contains 4 to 40 percent sorbital or sorbitanes, at least half of the weight of polyethylene glycol used is a polyethylene glycol having a mean molecular weight of 600, and the capsule filling comprises up to 20% by weight of glycerol and/or 1,2-propylene glycol.
WO 95/19579 (Dhabhar) teaches a process for solubilizing difficulty soluble pharmaceutical agents in a mixture of polyethylene glycol and propylene glycol by using a polyvinylpyrrolidone with a specific viscosity average molecular weight of from about 5,000 to about 25,000.
This invention provides orally administerable formulations for tricyclic vasopressin agonist compounds, or the pharmaceutically acceptable salts thereof, singularly or collectively optionally referred to herein as xe2x80x9cactive ingredientxe2x80x9d, which have the structure: 
wherein:
A, B, E, G are, independently, CH or nitrogen;
D is, independently, Cxe2x80x94W or nitrogen;
R1 is alkanoyl of 2 to 7 carbon atoms, a group selected from CN, COOH, CONH2, 
xe2x80x83or a moiety selected from the group: 
R2, R3 and R5 are, independently, hydrogen, straight chain alkyl of 1 to 6 carbon atoms, branched chain alkyl of 3 to 7 carbon atoms, cycloalkyl of 3 to 7 carbon atoms, or perfluoroalkyl of 1 to 6 carbons;
R4 is hydrogen, straight chain alkyl of 1 to 6 carbon atoms, branched chain alkyl of 3 to 7 carbon atoms, cycloalkyl of 3 to 7 carbon atoms, alkoxyalkyl of 2 to 7 carbon atoms, or an acyl substituent selected from the group consisting of alkanoyl of 2 to 7 carbon atoms, alkenoyl of 3 to 7 carbon atoms, cycloalkanoyl of 3 to 7 carbon atoms, aroyl, or arylalkanoyl;
and Y are, independently, hydrogen, straight chain alkyl of 1 to 6 carbon atoms, branched chain alkyl of 3 to 7 carbon atoms, cycloalkyl of 3 to 7 carbon atoms, perfluoroalkyl of 1 to 6 carbons, alkoxyalkyl of 2 to 7 carbon atoms, halogen (including chlorine, bromine, fluorine, and iodine), alkoxy of 1 to 6 carbons, hydroxy, CF3, or perfluoroalkyl of 2 to 6 carbons;
W is hydrogen, halogen (preferably chloro, bromo or iodo), alkyl, alkoxyalkyl of 2 to 7 carbons, hydroxyalkyl of 1 to 6 carbons, or CH2NR6R7;
R6 and R7 are, independently, hydrogen, straight chain alkyl of 1 to 6 carbon atoms, branched chain alkyl of 3 to 7 carbon atoms; or, taken together with the nitrogen atom of CH2NR6R7, R6 and R7 form a five or six membered ring optionally containing one or more additional heteroatoms such as, but not limited to, those of the group: 
R8 is a straight chain alkyl of 1 to 6 carbon atoms
R9 is independently hydrogen, trimethylsilyl or a straight chain alkyl of 1 to 6 carbon atoms;
or a pharmaceutically acceptable salt, ester or prodrug form thereof.
Among the more preferred active ingredient compounds of the formulations of this invention are those of the formula: 
wherein:
A and B are, independently, CH or nitrogen;
D is Cxe2x80x94W or nitrogen;
R1 is alkanoyl of 2 to 7 carbon atoms or a group selected from 
R2, R3 and R5 are, independently, hydrogen, straight chain alkyl of 1 to 6 carbon atoms, branched chain alkyl of 3 to 7 carbon atoms, cycloalkyl of 3 to 7 carbon atoms, or perfluoroalkyl of 1 to 6 carbons;
R4, X, Y, W, R6, R7 and R8 are as defined above;
or a pharmaceutically acceptable salt thereof.
For the compounds defined above and referred to herein, unless otherwise noted, aroyl groups include, for example, benzoyl, naphthoyl which can be substituted independently with one or more substituents from the group of hydrogen, halogen, cyano, straight chain alkyl of 1 to 6 carbon atoms, branched chain alkyl of 3 to 7 carbon atoms, alkoxy of 1 to 6 carbons, CF3, or phenyl.
Heteroaroyl groups herein refer to a carbonyl (radical ) directly bonded to a carbon atom of a five membered heterocyclic ring having one or two heteroatoms selected from nitrogen, oxygen, sulfur, for example 2-thienoyl. The heterocyclic ring of the heteroaroyl groups may also include, but are not limited to, groups in which the aryl portion is a furan, pyrrole, 2H-pyrrole, imidazole, pyrazole, isothiazole, isoxazole, thiophene, pyrazoline, imidazolidine or pyrazolidine group. The heteroaryl groups herein can be substituted independently with one or more substituents from the group of hydrogen, halogen, cyano, straight chain alkyl of 1 to 6 carbon atoms, or branched chain alkyl of 3 to 7 carbon atoms.
The arylalkanoyl groups herein refer to a carbonyl group or radical directly bonded to an alkyl group of 1 to 6 carbon atoms which is terminally substituted by an aryl group, for example, phenylacetic acid. The aryl group can be substituted independently with one or more substituents from the group of hydrogen, halogen, cyano, straight chain alkyl of 1 to 6 carbon atoms, branched chain alkyl of 3 to 7 carbon atoms, alkoxy of 1 to 6 carbons, CF3, or phenyl or substituted phenyl where the substituents are selected from halogen, cyano, straight chain alkyl of 1 to 6 carbon atoms, branched chain alkyl of 3 to 7 carbon atoms, alkoxy of 1 to 6 carbons, CF3.
The halogens referred to herein may be selected from fluorine, chlorine, bromine or iodine, unless otherwise specified.
It is understood by those practicing the art that the definition of the compounds of formula (I), when R1, R2, R3, R4, R5, R6, R7, X, or Y contain symmetric carbons, encompass all possible stereoisomers and mixtures thereof which possess the activity discussed below. In particular, it encompasses any optical isomers and diastereomers; as well as the racemic and resolved, enantiomerically pure R and S stereoisomers; as well as other mixtures of the R and S stereoisomers and pharmaceutically acceptable salts thereof, which possess the indicated activity. Optical isomers may be obtained in pure form by standard separation techniques. It is also understood that the definition of R1, R2, R3, R4, R5, R6, R7, X, or Y of the compounds of formula (I), encompasses all possible regioisomers, and mixtures thereof which possess the activity discussed below. Such regioisomers may be obtained pure by standard separation methods known to those skilled in the art.
Also among the preferred groups of active ingredients in the formulations of this invention are those in the subgroups:
a) compounds having the general formula: 
xe2x80x83wherein A, B, W, R1, R2, R3, R4, R5, R6, R7, R8, R9, X, and Y are as defined above;
b) compounds having the general formula: 
xe2x80x83wherein A, B, R1, R2, R3, R4, R5, R9, X, and Y, are as defined above; and
c) compounds having the general formula: 
xe2x80x83wherein A, B, R1, R2, R3, R4, R5, R9, X, and Y, are as defined above.
It is understood that subgroups a)-c), above, further include subgroups wherein:
A and B are, independently, CH or nitrogen;
R1 is alkanoyl of 2 to 7 carbon atoms or a group selected from 
R2, R3 and R5 are, independently, hydrogen, straight chain alkyl of 1 to 6 carbon atoms, branched chain alkyl of 3 to 7 carbon atoms, cycloalkyl of 3 to 7 carbon atom s, or perfluoroalkyl of 1 to 6 carbons; and
R4, X, Y, W, R6, R7 and R8 are as defined above;
or a pharmaceutically acceptable salt thereof.
Particularly preferred among the compounds of group a), above, are those in which W is H, A and B are each CH, and R1 is the group of alkanoyl of 2 to 7 carbon atoms or a group selected from the moieties (a), (b), (e), (f), (g), (h), (i) or (k), listed above.
The pharmaceutically acceptable salts include those derived from such organic and inorganic acids as: lactic, citric, acetic, tartaric, succinic, maleic, malonic, hydrochloric, hydrobromic, phosphoric, nitric, sulfuric, methanesulfonic, and similarly known acceptable acids.
Among the most preferred formulations of this invention are those described herein having as the active ingredient [2-Chloro-4-(3-methyl-pyrazol-1-yl)-phenyl-(5H, 11H)-pyrrolo[2,1-c][1,4]benzodiazepin-10-yl)-methanone, or pharmaceutically acceptable salts thereof, having the structure: 
The formulations of this invention are useful in methods for treating in humans or other mammals diseases, conditions or disorders in which vasopressin agonist activity is desired. These methods of treatment include those for diseases, conditions or disorders which make it desirable to release factor VIII and von Willebrand factor into the circulatory system, release tissue-type plasminogen activator (t-PA) in the blood circulation, or affect the renal conservation of water and urine concentration. Such methods of treatment include, but are not limited to, treatments for diabetes insipidus, nocturnal enuresis, nocturia, urinary incontinence, or bleeding and coagulation disorders in humans or other mammals, including hemophilia.
The methods herein for which these formulations are used include facilitation in humans or other mammals of temporary delay of urination, which may also be described as controlling or treating the inability to temporarily delay urination, whenever desirable. This method is understood to include treatments facilitating the temporary delay of urination which are separate from and not included in the treatment of the conditions known as nocturnal enuresis and nocturia.
In order to obtain consistency of administration, it is preferred that a composition of the invention is in the form of a unit dose. Suitable unit dose forms preferably include tablets or capsules, though one skilled in the art will understand semi-solids or gels of this invention are also readily made and useful. Such unit dose forms may contain from 0.1 to 1000 mg of a compound of the invention and preferably from 2 to 50 mg. Still further preferred unit dosage forms contain 1 to 25 mg, more preferably from 1 to 10 mg, of a compound of the present invention. The compounds of the present invention can be administered orally at a dose range of about 0.01 to 100 mg/kg or preferably at a dose range of 0.1 to 10 mg/kg. Such compositions may be administered from 1 to 6 times a day, more usually from 1 to 4 times a day.
The compositions of the invention may be formulated with other conventional carriers or excipients such as fillers, disintegrating agents, binders, lubricants, flavoring agents and the like.
The formulations of this invention comprise (by % w/w):
a) from about 1% to about 20% of active ingredient, or a pharmaceutically acceptable salt thereof, preferably from about 5% to about 16% of this active ingredient;
b) from about 1% to about 15% of a surfactant component, preferably from about 5% to about 10% of the surfactant component;
c) from about 50% to about 80% of a component of one or more olyethylene glycols (PEG), preferably from about 55% to about 70% of one or more olyethylene glycols; and
d) from about 1% to about 20%, preferably from about 5% to about 15% and more preferably between about 8% and about 12%, of a component of:
i) one or more sucrose fatty acid esters; or
ii) a polyvinylpyrrolidone (PVP) with a K value between about 15 and 90, preferably with a K value of from about 17 as defined in USP/NF; or
iii) a combination of one or more sucrose fatty acid esters and a PVP, as defined above.
The polyethylene glycol component may be comprised of one or more PEG polymers, preferably commercially available PEG polymers between PEG 200 and PEG 4,000, i.e. those PEG polymers having an average molecular weight between about 190 and about 4800. More preferred are PEG polymers between average molecular weights of from about 190 to about 3450, most preferably between about 400 and 1540. Among the preferred PEG polymers are PEG 400, having an average molecular weight between about 380 and about 420, and PEG 1,000, having an average molecular weight between about 950 and about 1050. The ratio of high and low molecular weight PEG species within the PEG component is preferably from about 2.5:1 to about 1:2.5, more preferably about 1:1. As an example, a preferred blend of PEG polymers within this invention would include a 1:1 blend of PEG 400 and PEG 1000. It may be preferable to choose a mixture of PEG components which will have a melting point at or near the physiological temperature of the mammal to receive the formulation. Mixtures of final components which have a viscosity range of from about 140 to about 1500 centipoise at 37xc2x0 C. may be preferred, more preferably a range of from 300 to about 800 centipoise at 37xc2x0 C.
The surfactants that may be used with the present formulations include, but not limited to, polysorbate 20 (polyoxyethylene 20 sorbitan monolaurate), Polysorbate 60, Polysorbate 40, polysorbate 80, Span 80 Sorbitan Oleate, a product of ICI Americas, Wilmington, Del., polysorbate 81, polysorbate 85, polysorbate 120, bile acids and salts defined by Martindale The Extra Pharmacopoeia Thirtieth Edition on page 1341-1342 such as Sodium taurocholates, Sodium deoxytaurocholates, Chenodeoxycholic acid, and ursodeoxycholic acid, and pluronic or poloxamers such as Pluronic F68, Pluronic L44, Pluronic L101, or combinations of one or more of the above. Polysorbate 80, by itself or in combination with one or more other surfactants, is preferred for use with this invention.
The sucrose fatty acid esters useful with this invention include those commercially available and art recognized esters useful for orally administered pharmaceutical compositions, including monoesters, diesters and triesters of sucrose, or mixtures or blends thereof. Specific examples of esters useful with this invention are sucrose monolaurate, sucrose monomyristate, sucrose monopalminate, sucrose monostearate, sucrose distearate, sucrose tristearate, sucrose trimyristate, and sucrose tripalmitate, or combinations thereof.
In addition to these components, other enhancing or protective antioxidants or preservatives may be added to the compositions of this invention, which may account for up to about 4% by weight of the formulation. Examples may include ascorbyl palmitate, benzyl alcohol, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc. Examples of these components in the present formulations would include BHA at a concentration from about 0.3% to about 2.5% (% w/w) and BHT at a concentration from about 0.005% to about 0.15% (% w/w), preferably with a mixture of BHA and BHT within these ranges.
A formulation of this invention utilizing one or more of these antioxidants or preservatives comprises:
a) from about 1% to about 20% of active ingredient, or a pharmaceutically acceptable salt thereof, preferably from about 5% to about 16% of this active ingredient;
b) from about 1% to about 18% of a surfactant component, preferably from about 5% to about 15% of the surfactant component, more preferably from about 8 to about 12% of the surfactant component;
c) from about 50% to about 80% of a component of one or more polyethylene glycols (PEG), preferably from about 55% to about 70% of one or more polyethylene glycols;
d) from about 1% to about 20%, preferably about 5% to about 15%, of one or more sucrose fatty acid esters or polyvinylpyrrolidone (PVP) with a K value between about 15 and 90, preferably with a K value of from about 17 as defined in USP/NF; and
e) from about 0.1% to about 4% of one or more preservatives or antioxidants, for example from about 0.3% to about 2.5% (% w/w) BHA and/or from about 0.005% to about 0.15% (% w/w) BHT.
One preferred embodiment of this invention provides a pharmaceutical formulation comprising:
a) from about 5% to about 16% of active ingredient;
b) from about 5% to about 10% of a surfactant component;
c) a component of from about 55% to about 70% of one or more polyethylene glycols;
d) from about 5% to about 15% of polyvinylpyrrolidone (PVP) with a K value between about 15 and 90, preferably with a K value of from about 17 as defined in USP/NF; and
e) from about 0.3% to about 2.5% (% w/w) BHA and from about 0.005% to about 0.15% (% w/w) BHT.
Preferably, the formulations of this invention are enclosed in a sealed enclosure after manufacture, such as soft or hard gelatin capsules. The formulations of this invention may be created as a liquid or semi-liquid formulation and introduced into a capsule. Similarly, using an acceptable range of components and/or temperatures, the formulation may be made as a gel or solid prior to encapsulation.
The active compounds useful in the formulations of the present invention may be prepared according to one of the general processes outlined below.
As shown in Scheme I, a tricyclic benzodiazepine of formula (1) is treated with an appropriately substituted acetylaroyl (heteroaroyl) halide, preferably an aroyl (heteroaroyl) chloride of formula (2) in the presence of a base such as pyridine or a trialkylamine such as triethylamine, in an aprotic organic solvent such as dichloromethane or tetrahydrofuran at temperatures from xe2x88x9240xc2x0 C. to 50xc2x0 C. to yield the acylated derivative of formula (3). Treatment of (3) with a dialkylamide dialkyl acetal of formula (4) in an aprotic organic solvent such as dichloromethane at temperatures ranging from 0xc2x0 C. to the reflux temperature of the solvent yields the enone of formula (5) according to the procedure of Lin et al., J. Het. Chem., 14, 345 (1977). Treatment of (5) with hydroxylamine or a substituted hydrazine of formula (6) in acetic acid at temperatures ranging from ambient to the reflux temperature of the solvent yields the target compounds of formula (I) wherein A, B, D, E, G, X, Y, R2 and R4 are as defined above, and R1 is an heterocyclic moiety selected from the (f), (g), or (j) group of heterocycles defined above. 
The preferred substituted acetylaroyl (heteroaroyl) chlorides of formula (2) of Scheme I are conveniently prepared by treating the corresponding carboxylic acids with thionyl chloride at temperatures ranging from ambient to the reflux temperature of the solvent, or with oxalyl chloride in an aprotic solvent such as dichloromethane or tetrahydrofuran in the presence of a catalytic amount of dimethylformamide at temperatures ranging from 0xc2x0 C. to 40xc2x0 C.
The preferred dialkylamide dialkylacetals are either available commercially, or are known in the literature, or can be conveniently prepared according to procedures analogous to those in the literature. Kantlehner, W. Chem. Ber. 105, 1340 (1972).
The preferred tricyclic benzodiazepines of formula (1) are a 10,11-dihydro-5H-pyrrolo[2,1-c][1,4]benzodiazepine (Albright et al., U.S. Pat. No. 5,536,718, issued Jul. 16, 1996), a 10,11-dihydro-5H-pyrazole[5,1-c][1,4]benzodiazepine, Cecchi, L. et. al., J. Het. Chem., 20, 871 (1983). and 10,11-dihydro-5H-tetrazole[5,1-c][1,4]benzodiazepine, Klaubert, D. H., J. Het. Chem., 22, 333 (1985). 
An alternate process for the preparation of intermediates of formula (3) is illustrated in the following Scheme II. 
Thus, a tricyclic benzodiazepine of formula (1) is treated with an appropriately substituted bromo aroyl (heteroaroyl) halide, preferably an aroyl (heteroaroyl) chloride of formula (8) in the presence of an organic base such as pyridine or a trialkylamine such as triethylamine in an aprotic organic solvent such as dichloromethane or tetrahydrofuran at temperatures from xe2x88x9240xc2x0 C. to 50xc2x0 C. to yield the acylated intermediate of formula (9). The intermediate (9) is subsequently coupled with a mono substituted terminal acetylene such as trimethylsilyl or a straight chain alkyl of 1 to 6 carbon atoms, in the presence of pyridine and a catalyst such as bis(triphenylphosphine) palladium (II) chloride and copper (I) iodide in an organic base such as triethylamine as the solvent, in a sealed pressure tube at temperatures ranging from ambient to 100xc2x0 C. essentially according to the procedure of Martinez et al., J. Med. Chem., 35, 620 (1992). The resulting acetylene intermediate of formula (10) is then hydrated by treatment with 1% sulfuric acid in an aprotic organic solvent such as tetrahydrofuran saturated with mercury (II) sulfate at ambient temperature essentially according to the procedure of Reed et al., J. Org. Chem., 52, 3491 (1987) to provide the desired acyl compound of formula (3) wherein A, B, D, E, G, X, and Y, are as defined above and R9 is hydrogen or a straight chain alkyl of 1 to 6 carbon atoms. Alternatively, compound 9 where R9 is trimethyl is treated with tetrabutylbutylammonium fluoride in an ether solvent such as tetrahydrofuran to afford compound (10) where R9 is hydrogen.
The preferred acylating agents of formula (8) of Scheme II are conveniently prepared by treating an appropriately substituted aryl (heteroaryl) carboxylic acid of formula (7) with thionyl chloride at temperatures ranging from ambient to the reflux temperature of the solvent, or with oxalyl chloride in an aprotic organic solvent such as dichloromethane or tetrahydrofuran in the presence of a catalytic amount of dimethylformamide at temperatures ranging from 0xc2x0 C. to 40xc2x0 C.
The protected acetylene intermediates of Scheme II are either available commercially, or are known in the art, or can be readily prepared by procedures analogous to those in the art.
As shown in Scheme III, the intermediate acetyl compounds (3) of Scheme I can be prepared also by the Stille coupling of a bromo aryl (heteroaryl) compound of formula (9) of Scheme II with a (xcex1-ethoxyvinyl)trialkyltin, preferably a (xcex1-ethoxyvinyl)tributylltin, in the presence of a catalytic amount of bis(triphenylphosphine) palladium(II) chloride in an aprotic organic solvent such as toluene at temperatures ranging from ambient to the reflux temperature of the solvent, essentially according to the procedure of Kosugi et al., Bull. Chem. Soc. Jpn., 60, 767 (1987). 
The preparation of the acetyl compound (3) can also be accomplished via a palladium-catalyzed arylation of a vinyl alkyl ether such as vinyl butylether, with the aryl halide intermediate of formula (9) according to the procedure of Cabri et al., Tetrahedron Lett., 32, 1753 (1991).
The (a-alkoxyvinyl)trialkyltin intermediates of Scheme III are either available commercially, or are known in the art, or can be readily prepared by procedures analogous to those in the art.
In the case where R4 in Scheme I is hydrogen, the heterocyclic nitrogen can be alkylated or acylated according to the reactions outlined in Scheme IV. 
Thus, the pyrazole compound of formula (I, R4 is H) is alkylated by treatment with a strong base such as sodium or potassium hydride and an alkylating agent such as an alkyl halide, preferably an alkyl chloride (bromide or iodide) in an aprotic organic solvent such as dimethylformamide or tetrahydrofuran at temperatures ranging from 0xc2x0 C. to 80xc2x0 C. to yield compound (I, R1=(f) or (g)) wherein A, B, D, E, G, X, Y, and R2 are as defined above, and R4 is an alkyl or acyl moiety. Alternatively, compound (I) is acylated by treatment with a carboxylic acid halide, preferably a chloride, or a carboxylic acid anhydride in the presence of an amine base such as pyridine or a trialkylamine, preferably triethylamine, in an aprotic organic solvent such as dichloromethane or tetrahydrofuran with no additional solvent when pyridine is used as the base, at temperatures ranging from xe2x88x9240xc2x0 C. to ambient to yield compound (I) wherein A, B, D, E, G, X, Y and R2 are as defined above, and R4 is an alkyl or acyl moiety. The alkylation or acylation of a compound of formula (I, R4 is H) leads to a mixture of regioisomers wherein R2 is hydrogen and R1 is an heterocyclic moiety selected either from the (f) or (g) group of heterocycles defined above and illustrated below, respectively. 
The compounds of general formula (I) of Scheme I wherein A and B are carbon, R2 is H, and R1 is an heterocyclic moiety selected from the (g) group of heterocycles defined above, can be prepared according to the general process outlined in Scheme V. 
Thus, an appropriately substituted haloaryl (heteroaryl) carboxylic acid ester, preferably a bromo (or iodo) methylester of formula (11) is coupled with a dialkylamino propyne, preferably 1-dimethylamino propyne, in the presence of a catalyst such as bis(triphenylphosphine) palladium(II) chloride and copper(I) iodide in an organic base such as triethylamine as the solvent and at temperatures ranging from ambient to 80xc2x0 C. essentially according to the procedures of Alami et al., Tetrahedron Lett. 34, 6403 (1993), and of Sanogashira et al., Tetrahedron Lett., 4467 (1975) to provide the substituted acetylene intermediate of general formula (12). The intermediate (12) is subsequently converted into its N-oxide by treatment with an oxidizing agent using any of a number of standard oxidative procedures (Albini, A., Synthesis, 263 (1993) or with dioxirane reagents (Murray, R. W., Chem. Rev., 1187 (1989), in an aprotic organic solvent such as dichloromethane at temperatures below ambient. The intermediate N-oxide is not isolated but is rearranged in situ to an enone of general formula (13) by treatment with, preferably with heating, a hydroxylic solvent, including any solvent or combination of solvents composed of or containing water, any C1-C8 straight chain or branched chain alkyl alcohol, ethylene glycol, polyethylene glycol, 1,2-propylene diol, polypropylene glycol, glycerol, 2-methoxyethanol, 2-ethoxyethanol, 2,2,2-trifluoroethanol, benzyl alcohol, phenol, or any equivalent solvent that contains one or more free hydroxyl (xe2x80x94OH) substituent(s) that is known to those skilled in the art.
Solvent systems containing one or more cosolvents, along with one or more solvents may also be used for this process of rearranging the N-oxide to the desired enaminone. The cosolvents referred to herein may be defined as a diluent of the main solvent(s) and can be selected from: hydrocarbons such as pentane, hexane or heptane; aromatic hydrocarbon such as benzene, toluene or xylene; ethers such as diethyl ether, tetrahydrofuran, dioxane or dimethoxy ethane; chlorinated hydrocarbons such as dichloromethane, chloroform, dichloroethane, or tetrachloroethane; or other common solvents such as ethyl acetate, N,N-dimethylformamide, N,N-dimethylacetamide, acetonitrile, dimethylsulfoxide, acetone, or the like.
The conversion of the amine N-oxide into an enaminone may be accomplished by introducing the amine N-oxide into a suitable hydroxylic solvent, preferably with stirring, at or between about room or ambient temperature and about the reflux temperature of the solvent. In other instances the introduction of the amine N-oxide to a hydroxylic solvent, preferably with stirring, may be accomplished in the presence of an acceptable catalyst, such as a palladium(II) catalyst or a copper (I) catalyst, at or between room temperature and the reflux temperature of the solvent.
This procedure provides a novel synthesis of enaminone compounds from propargylic amines or their N-oxides in hydroxylic solvents, which influence the ultimate outcome of the reaction. This new method of enaminone synthesis provides a convenient alternative to existing methods and further extends the range of starting materials that can be converted into enaminone products.
Although the precise mechanism by which a propargylic amine N-oxide is converted into an enaminone product has not been rigorously determined, it likely resembles two known processes; the thermal [2,3]-sigmatropic rearrangement of propargylic amine N-oxides (Craig, et. al., Tetrahedron Lett., 4025, 1979; Hallstrom, et. al., Tetrahedron Lett., 667, 1980; Khuthier, A-H, et. al., J. Chem. Soc. Chem. Commun., 9, 1979) and the conversion of certain isoxazoles into enaminones (Liguori, et. al., Tetrahedron, 44, 1255, 1988).
Treatment of (13) with a substituted hydrazine (6) in acetic acid at temperatures ranging from ambient to reflux leads to a mixture of regioisomeric compounds of general formulas (14) and (15) in a variable ratio. The major isomer of formula (14) is separated by means of chromatography and/or crystallization and subsequently hydrolyzed to the desired carboxylic acid of formula (16).
The intermediate (16) is then converted into an acylating species, preferably an acid chloride (bromide or iodide) or a mixed anhydride of formula (17) by procedures analogous to those described hereinbefore. The acylating agent (17) is then used to acylate a tricyclic benzodiazepine of formula (1) by any of the procedures described hereinbefore to yield the desired compound of formula (I), wherein A, B are CH and D, E, G, X, Y, and R4 are as defined above, R2 is hydrogen and R1 is a heterocyclic moiety selected from the (g) group of heterocycles illustrated below. 
Likewise, treatment of (13) with an unsubstituted hydrazine (6, R4 is H) in acetic acid at temperatures ranging from ambient to the reflux temperature of the solvent yields the intermediate pyrazole ester of formula (18). In this case the heterocyclic nitrogen can be alkylated or acylated as shown in Scheme VI to provide compounds of formula (I) wherein R2 is hydrogen, and R1 is an heterocyclic moiety selected from the (f) group of heterocycles defined above. 
Thus, the intermediate ester of formula (18) is alkylated by treatment with a strong base such as sodium or potassium hydride and an alkylating agent such as an alkyl halide, preferably an alkyl chloride (bromide or iodide), in an aprotic solvent such as dimethylformamide or tetrahydrofuran at temperatures ranging from 0xc2x0 C. to 80xc2x0 C. to yield a mixture of regioisomers of formulas (14) and (15) in a variable ratio. The major regioisomer of formula (15) is separated by chromatography and/or crystallization and subsequently hydrolyzed to the desired carboxylic acid of formula (19), which is then converted into an acylating agent, preferably an acyl chloride or a mixed anhydride by procedures analogous to those described hereinbefore. The acylating species of formula (20) is then used to acylate a tricyclic benzodiazepine of formula (1) to yield the desired compound of formula (I), wherein A, B, D, E, G, X, Y, and R4 are as defined above, R2 is hydrogen, and R1 is a heterocyclic moiety selected from the (f) group of heterocycles defined above. 
Compounds of general formula (I) wherein R1 is an heterocyclic moiety selected from the (h) group of R1 heterocycles defined above, can be prepared as outlined in Scheme VII. 
An appropriately substituted malondialdehyde of formula (21) is treated first with a hydrazine in acetic acid at temperatures ranging from ambient to the reflux temperature of the solvent and the intermediate pyrazole is then oxidized with potassium permanganate in a basic aqueous solution at temperatures ranging from ambient to the reflux temperature of the solvent to yield a carboxylic acid intermediate of formula (22). The acid (22) is converted into an acylating agent, preferably an acid chloride (bromide or iodide) or a mixed anhydride by procedures analogous to those described hereinbefore. The acylating agent of formula (23) is finally reacted with a tricyclic benzodiazepine of formula (1) to yield compounds of general formula (I) wherein A, B, D, E, G, X, Y, and R4 are as defined above, and R1 is a heterocyclic moiety selected from the (h) group of heterocycles defined above. 
In the case where R4 in Scheme VII is hydrogen, the heterocyclic nitrogen can be alkylated or acylated according to the procedures outlined hereinbefore.
The preferred malondialdehydes of formula (21) and the hydrazines of Scheme VII are either available commercially, or are known in the art, or can be readily prepared by procedures analogous to those in the literature for known compounds, such as those of Knorr et al., J. Org. Chem., 49, 1288 (1984) and Coppola et al., J. Het. Chem., 11, 51 (1974).
An alternative preparation of the intermediate carboxylic acids of formula (22) of Scheme VII wherein Y is as defined above and R4 is other than hydrogen, is outlined in Scheme VIII. 
The organotin reagent of formula (25) is reacted in a Stille coupling reaction with an appropriately substituted aryl (heteroaryl) halide, preferably a bromide or iodide of formula (28) in the presence of a catalyst such as tetrakis(triphenylphosphine)palladium (0) and copper (I) iodide in an organic aprotic solvent such as dimethylformamide at temperatures ranging from ambient to 150xc2x0 C., essentially according to procedures analogous to those found in Farina et al., J. Org. Chem., 59, 5905 (1994). Basic hydrolysis of the resulting ester of formula (26) with sodium or lithium hydroxide in aqueous alcohol or tetrahydrofuran at temperatures ranging from ambient to the reflux temperature of the solvent yields the desired carboxylic acids of formula (22).
In turn, the organotin reagents of formula (25) wherein the R groups are preferably alkyl groups, are conveniently prepared by metallation of a 4-bromo N-alkylpyrazole of formula (24) with a trialkyltin halide, preferably a tributyltin chloride (or bromide) in the presence of a metallating agent such as an alkyllithium such as n-butyl lithium, sec-butyl lithium, or tert-butyllithium in an aprotic organic solvent such as diethylether at temperatures ranging from xe2x88x9240xc2x0 C. to ambient according to procedures analogous to those found in Martina et al., Synthesis, 8, 613 (1991).
The preferred N-alkyl substituted 4-bromo pyrazoles of formula (24) are conveniently prepared from 4-bromo pyrazole by alkylation with an alkyl halide, preferably an alkyl chloride (bromide or iodide) in the presence of a strong base such as lithium, sodium or potassium hydride in an aprotic organic solvent such as dimethylformamide or tetrahydrofuran at temperatures ranging from 0xc2x0 C. to 80xc2x0 C. Alternatively, alkylation of 4-bromopyrazole can be carried out with an alkylating agent mentioned above, and a strong alkaline base such as lithium, sodium or potassium hydroxide in the presence of a phase transfer catalyst (Jones, R. A. Aldrichimica ACTA, 9(3), 35, 1976) such as benzyldimethyltetradecylammonium chloride, or benzyltrimethylammonium chloride.
The preferred aryl (heteroaryl) iodides of formula (28) are conveniently prepared by diazotization of the corresponding substituted anilines of formula (27) followed by reaction of the corresponding diazonium salt with iodine and potassium iodide in aqueous acidic medium essentially according to the procedures of Street et al., J. Med. Chem., 36, 1529 (1993) and of Coffen et al., J. Org. Chem., 49, 296 (1984).
An alternative preparation of compounds of general formula (I) is outlined in Scheme IX. 
A tricyclic benzodiazepine of formula (1) is treated with an appropriately substituted haloaroyl (heteroaroyl) halide, preferably a fluoro aroyl or a fluoro (or chloro) heteroaroyl chloride of formula (29), in the presence of a base such as triethylamine or diisopropylethylamine in an aprotic organic solvent such as dichloromethane or tetrahydrofuran at temperatures from xe2x88x9240xc2x0 C. to the reflux temperature of the solvent to yield the acylated derivative (30).
Alternatively, the acylating species can be a mixed anhydride of the carboxylic acid described above, such as that prepared by reaction 2,4,6-trichlorobenzoyl chloride in a solvent such as dichloromethane according to the procedure of Inanaga et al., Bull. Chem. Soc. Jpn, 52, 1989 (1979). Treatment of said mixed anhydride of general formula (29) with the tricyclic benzodiazepine of formula (1) in a solvent such as dichloromethane and in the presence of an organic base such as 4-dimethylaminopyridine at temperatures ranging from 0xc2x0 C. to the reflux temperature of the solvent, yields the intermediate acylated derivative (30) of Scheme IX.
A compound of formula (30) is then treated with the lithium, sodium or potassium salt of an appropriately substituted heterocycle of formula (31) in a polar aprotic organic solvent such as dimethylformamide or tetrahydrofuran at temperatures ranging from ambient to the reflux temperature of the solvent to yield a compound of general formula (I), wherein A, B, D, E, G, X, Y, R2, R3, and R5 are as defined above, and R1 is a heterocyclic moiety selected from the group consisting of (a), (b), (c), (d), (l), (n) or (o) defined above. 
The condensation of the intermediate of formula (30) with the intermediate salt of formula (31) leads to a variable ratio of regioisomers of general formula (I) which are separated by means of chromatography and/or crystallization.
The preferred substituted fluoro aroyl and fluoro (or chloro) heteroaroyl chlorides of formula (29) are either available commercially, or are known in the art, or can be readily prepared by procedures analogous to those in the literature for the known compounds.
The lithium, sodium or potassium salts of the heterocycles of formula (31) are prepared by treatment of said heterocycle with a strong base such as lithium hydride, sodium hydride, potassium hydride or a metal alkoxide at temperatures ranging from xe2x88x9240xc2x0 C. to ambient in an aprotic organic solvent such as dimethylformamide or tetrahydrofuran.
Alternatively, the compounds of general formula (I) described in Scheme IX can be prepared according to the process outlined in Scheme X. 
Thus, an appropriately substituted fluoroaryl or fluoro (or chloro)heteroaryl carboxylic acid of formula (32) is esterified using methods known in the art such as treatment with oxalyl chloride (or thionyl chloride) in an alcohol solvent such as methanol, in the presence of a catalytic amount of dimethylformamide; or by condensation with methanol in the presence of an acid catalyst such as para-toluenesulfonic acid at temperatures ranging from ambient to reflux.
The resulting ester of formula (33) is reacted with the lithium, sodium or potassium salt of an appropriately substituted heterocycle of formula (31) in a polar aprotic organic solvent such as dimethylformamide at temperatures ranging from ambient to 150xc2x0 C., to yield an intermediate ester of formula (34). The condensation of (33) with (31) leads to a variable ratio of regioisomers of formula (34) which are separated by means of chromatography and/or crystallization.
Subsequent hydrolysis of the intermediate ester of formula (34) with an aqueous base such as lithium, sodium or lithium hydroxide in methanol or tetrahydrofuran affords the carboxylic acid of formula (35).
The intermediate carboxylic acid (35) is then converted into an acylating agent preferably an acid chloride or a mixed anhydride of general formula (36) using any of the procedures described hereinbefore.
Subsequent reaction of the tricyclic benzodiazepine of formula (1) with the intermediate acylating agent of formula (36) according to any of the procedures described hereinbefore yields the desired compounds of formula (I) of Scheme IX.
Alternatively, the substituted carboxylic acids of formula (35) described in Scheme X can be prepared according to the process outlined in Scheme XI. 
Thus, a fluoro aryl or fluoro (chloro)heteroaryl nitrile of formula (37) is reacted with the lithium, sodium or potassium salt of a substituted heterocycle of formula (31) in an apolar aprotic solvent such as dimethylformamide at temperatures ranging from ambient to 150xc2x0 C., to yield an intermediate of general formula (38). The reaction of (37) with (31) leads to a variable ratio of regioisomers of formula (38) which are separated by means of chromatography and/or crystallization. Hydrolysis of the intermediate nitrites of formula (38, Yxe2x89xa0CF3) is preferentially carried out with an inorganic acid such as sulfuric acid at temperatures ranging from ambient to 60xc2x0 C.
Alternatively, hydrolysis of the nitrile (38) can be carried out by heating in ethanol in the presence of a strong alkaline base such as sodium hydroxide with or without a phase transfer catalyst (Jones, R. A. Aldrichimica Acta, 9(3), 35, 1976,) such as benzyldimethyltetradecyl ammonium chloride.
The resulting carboxylic acids of formula (35) are then converted into the desired compounds of formula (I) of Scheme IX by procedures analogous to those described hereinbefore.
Alternatively, the substituted carboxylic acids of formula (35) of Scheme X can be prepared according to the process outlined in Scheme XII by sequential treatment of a nitrile of formula (38) wherein A and B are CH and where R1 is not alkanoyl of 2 to 7 carbons, alkynyl, (b) or (d), with basic hydrogen peroxide in dimethylsulfoxide essentially according to the procedure of Katritzky et al., Synthesis, 949 (1989), followed by hydrolysis of the resulting amide of formula (38) preferably by treatment with dilute sulfuric acid and sodium nitrite according to the procedure of Hanes et al., Tetrahedron, 51, 7403 (1995). 
Where R1 is not (b) or (d)
A preferred process for the preparation of the intermediate substituted carboxylic acids of formula (35) of Scheme X wherein R1 is a heterocyclic moiety selected from the (a) group of R1 heterocycles defined above, is outlined in Scheme XIII. 
Diazotization of an appropriately substituted aniline of formula (40) followed by reduction of the resulting diazonium salt of formula (41) with tin (II) chloride in concentrated hydrochloric acid according to the procedure of Street et al., J. Med. Chem., 36, 1529 (1993) provides the intermediate hydrazine hydrochloride salt of formula (42). Subsequent condensation of (42) with an aldehyde derivative of formula (47), wherein R2 is as defined above, R3 and R5 is H, and P is dialkylacetal) such as acetylacetaldehyde dimethyl acetal, or a ketone of formula (47), wherein R2, R3 and R5 are as defined above, and P is xe2x95x90O or (O-alkyl)2 in a solvent such as aqueous methanol at temperatures ranging from ambient to 100xc2x0 C. provides after crystallization, the desired intermediate ester of formula (34, R1 is (a) and R5 is H), which is then converted to the compound of formula (I) as outlined in Scheme X above. 
When Y is OCH3 the compounds of general formula (I) of Scheme I can be conveniently demethylated as outlined in Scheme XIV. 
Thus, the reaction of compound (I) wherein Y is OCH3 with boron tribromide in an organic solvent, such as dichloromethane, yields the corresponding phenol of formula (I) wherein Y is OH, and A, B, D, E, G, X, R2 and R3 are as defined above and R1 is an heterocyclic moiety selected from the group (a) of heterocycles defined above and illustrated below. 
Compounds in which R1 contains three heteroatoms are prepared according to Scheme XV. 
Thus, a tricyclic benzodiazepine of formula (1) is treated with an appropriately substituted cyano aroyl (heteroaroyl) halide, preferably an aroyl (heteroaroyl) chloride of formula (43) in the presence of a base in an aprotic organic solvent such as dichloromethane or tetrahydrofuran at temperatures ranging from xe2x88x9240xc2x0 C. to 80xc2x0 C. to yield an intermediate nitrile of formula (46, Scheme XVI) which in turn, is hydrolyzed to an amide intermediate of general formula (44) with an inorganic acid such as sulfuric acid at ambient temperature to 50xc2x0 C. Treatment of the amide (44) with a dialkyl amide dialkyl acetal of formula (4) in an aprotic organic solvent such as dichloromethane or tetrahydrofuran at temperatures ranging from 0xc2x0 C. to 80xc2x0 C. yields the intermediate of formula (45). Treatment of (45) with hydroxylamine or a hydrazine of formula (6) in acetic acid at temperatures ranging from ambient to reflux yields the desired target compounds of formula (I) wherein A, B, D, E, G, X, Y, R2 and R4 are as defined above, and R1 is an heterocyclic moiety selected from the (e), (i) and (k) group of heterocycles defined above. 
Another preferred process for the preparation of the intermediate amide of formula (44), see Scheme XV, wherein A and B are CH and D is not CH is outlined in Scheme XVI and consists of treating a nitrile of formula (46) with basic hydrogen peroxide in dimethylsulfoxide essentially according to the procedure of Katritzky et al., Synthesis, 949 (1989). 
The preferred process to prepare compounds of general formula (I) in which R1 contains four heteroatoms and R4 is hydrogen is outlined in Scheme XVII. 
Treatment of the nitrile intermediate of formula (46) of Scheme XVI with sodium azide and ammonium chloride in an aprotic organic solvent such as dimethylformamide at temperatures ranging from ambient to the reflux temperature of the solvent yields the desired compounds of formula (I) wherein A, B, D, E, G, X, and Y, are as defined above, R4 is hydrogen and R1 is an heterocyclic moiety selected from the group (m) of heterocycles defined above. 
The compounds of general formula (I) wherein D is CW and W is hydrogen, can undergo Mannich condensation as shown in Scheme XVIII. 
Thus, reaction of compounds of formula (I, D is CH) with either aqueous formaldehyde or paraformaldehyde , a substituted amine of formula (47), and glacial acetic acid in an alcohol solvent such as methanol at temperatures ranging from ambient to reflux yields the corresponding Mannich bases of general formula (I), wherein A, B, E, G, X, Y, R2, R3, R5, R6 and R7 are as defined above; D is CW; W is a dialkylaminoalkyl residue preferably a dimethylaminomethyl residue, and R1 is an heterocyclic moiety selected from the (a), (c), (e), (f), (g), (h), (i), (j), (k), (l), (m), (n) and (o) group of heterocycles defined above.
Likewise, the compounds of general formula (I) wherein D is CH can undergo halogenation as shown in Scheme XIX. 
Thus, reaction of (I, D is CH) with a N-halosuccinimide such as N-chloro (bromo or iodo)succinimide in a polar aprotic organic solvent such as dichloromethane at temperatures ranging from xe2x88x9280xc2x0 C. to ambient yields the corresponding halogenated derivatives of general formula (I), wherein A, B, E, G, X, R2, R3 and R5 are as defined above, D is CW, W is a halogen such as chlorine (bromine or iodine), and R1 is an heterocyclic moiety selected from the (a), (c), (e), (f), (g), (h), (i), (j), (k), (l), (m), (n) and (o) group of heterocycles defined above.
The subject compounds of the present invention were tested for biological activity according to the following procedures.
Male or female normotensive Sprague-Dawley rats (Charles River Laboratories, Inc., Kingston, N.Y.) of 350-500 g body weight were supplied with standard rodent diet (Purina Rodent Lab. Chow 5001) and water ad libitum. On the day of test, rats were placed individually into metabolic cages equipped with devices to separate the feces from the urine and containers for collection of urine. Test compound or reference agent was given at an oral dose of 10 mg/kg in a volume of 10 ml/kg. The vehicle used was 20% dimethylsulfoxide (DMSO) in 2.5% preboiled corn starch. Thirty minutes after dosing the test compound, rats were gavaged with water at 30 ml/kg into the stomach using a feeding needle. During the test, rats were not provided with water or food. Urine was collected for four hours after dosing of the test compound. At the end of four hours, urine volume was measured. Urinary osmolality was determined using a Fiske One-Ten Osmometer (Fiske Associates, Norwood, Mass., 02062) or an Advanced CRYOMATIC Osmometer, Model 3C.2 (Advanced Instruments, Norwood, Mass.). Determinations of Na+, K+ and Clxe2x88x92 ion were carried out using ion specific electrodes in a Beckman SYNCHRON EL-ISE Electrolyte System analyzer. The urinary osmolality should increase proportionally. In the screening test, two rats were used for each compound. If the difference in the urine volume of the two rats was greater than 50%, a third rat was used.
Male or female homozygous Brattleboro rats (Harlan Sprague Dawley, Inc., Indianapolis, Ind.) of 250-350 g body weight were supplied with standard rodent diet (Purina Rodent Lab. Chow 5001) and water ad libitum. On the day of test, rats were placed individually into metabolic cages equipped with devices to separate the feces from the urine and containers for collection of urine. Test compound or reference agent was given at an oral dose of 1 to 10 mg/kg in a volume of 10 ml/kg. The vehicle used was 20% dimethylsulfoxide (DMSO) in 2.5% preboiled corn starch. During the test, rats were provided with water ad libitum. Urine was collected for six hours after dosing of the test compound. At the end of six hours, urine volume was measured. Urinary osmolality was determined using a Fiske One-Ten Osmometer (Fiske Associates, Norwood, Mass., 02062) or an Advanced CRYOMATIC Osmometer, Model 3C2 (Advanced Instruments, Norwood, Mass.). Determinations of Na+, K+ and Clxe2x88x92 ion were carried out using ion specific electrodes in a Beckman SYNCHRON EL-ISE Electrolyte System analyzer. This animal model was mainly used for evaluation of potency and duration of action of the active compounds. The results of this study are shown in Table I.